Hygiena MicroSnap MS2-EB User manual

INS0193 JAN 2015 Rev A
MicroSnap Enhanced EB Broth –9ml
Directions for use with MicroSnap EB Detection Device (MS2-EB)
Matrix: Liquid Milk Products
Part No: MS1-EB-BROTH-9ML (100)
Description/Intended Use:
Hygiena’s Enhanced EB Broth is a ready-to-use device compatible with
MicroSnap EB (MS2-EB), MicroSnap Coliform (MS2-COLIFORM) and
MicroSnap E. coli (MS2-ECOLI) detection devices.
Instructions in this insert are for MicroSnap EB (MS2-EB) Detection
Devices with liquid milk samples only.
Enhanced EB Broth contains 9ml of unique liquid medium designed to grow
aerobic and facultative microorganisms while enhancing production of
biomarkers and specific enzymes diagnostic of Enterobacteriaceae,and
reducing sample interferences. The broth has been primarily produced for
liquid milk products. For use with other product samples or incubation times
and temperatures other than 37 ± 0.5°C, please contact Hygiena for
guidance.
Required Materials (Not Provided):
EnSURE luminometer (Part No. ENSURE)
MicroSnap EB Detection Device (MS2-EB)
Dry heating block incubator at 37 ± 0.5°C
Pipettes for transferring 1ml and 0.1ml volumes
Vortex (optional)
Directions:
Instructional Video: www.youtube.com/HygienaTV
Step 1: Enrichment
Enrichment procedure is described below and is alsoshown in Step 1
diagrams.
Visually inspect liquid in tube prior to use. Liquid should be clear and light
straw color, not turbid or cloudy. Use permanent marker to identify sample
on tube label.
1. Allow milk sample and Enhanced EB Broth vial to equilibrate to room
temperature (10 minutes at 22 –26 °C). Unscrew Enhanced EB Broth
cap and aseptically add 1ml liquid milk sample.
2. Replace cap and tighten to secure.
3. Mix vial contents by hand-shaking or vortexing for 10 seconds.
Incubate at 37 ± 0.5°C for 6 to 8 hours, depending on sensitivity
required. Refer to Table 1 below.
Table 1: Incubation Time & Dynamic Range
Incubation Time
CFU Range
Results
6 hrs
500-250,000
Enumeration
7 hrs
50-50,000
Enumeration
8 hrs
<5-5000
Presence/Absence
Step 2: Detection
Detection procedure is described below and is also shown in Step 2
diagrams.
Turn on luminometer. If locations have been programmed, select location to
be tested.
1. Allow MicroSnap EB Detection Device (Part # MS2-EB) to equilibrate
to room temperature (10 minutes at 22 –26°C). Shake test device by
either tapping on palm of hand 5 times, or forcefully flicking in a
downward motion once. This will bring extractant liquid to bottom of
tube.
2. Remove Enhanced EB Broth tube from incubator andmix by manually
shaking or vortexing for 10 seconds to disperse sample.
3. Open Detection Device by twisting and pulling to remove bulb. Set
aside. Using pipette, aseptically transfer 0.1ml of enriched sample
directly into Detection Device tube.
4. Reassemble Detection Device to original state.
5. Activate Detection Device by holding swab tube firmly and using
thumb and forefinger to break Snap-Valve by bending bulb forward
and backward. Squeeze bulb 3 times to release all liquid to bottom of
swab tube.
6. Shake gently to mix.
7. Immediately insert whole device into luminometer; close lid and
holding unit upright, press “OK” button to initiate measurement.
Results will appear after 15 second count down.
8. Result will be displayed in RLU (relative light units). Set RLU
thresholds on instrument to correspond with required CFU limits.
Refer to “Interpretation of Results” below for correlation.
Interpretation of Results:
Results are displayed as relative light units (RLU). RLU output is
proportional to the starting inoculums and corresponding bacteria
equivalent numbers (expressed as colony forming units, CFU). Table 2
shows equivalent colony forming unit (CFU) values to RLU. This will give an
estimation of the Enterobacteriaceae CFU/mL from the original milk sample.
Table 2: Average relationship between CFU and MicroSnap EB RLU at 6, 7
and 8 hours incubation of 1mL of whole milk naturally contaminated with
organisms at various levels (n=25 per level) incubated at 37 ± 0.5°C
mean CFU/mL
EnSURE RLU
6 hours
7 hours
8 hours
<5
NA
NA
<5
10
NA
NA
<10
25
NA
NA
<25
50
NA
<10
<50
100
NA
<20
<100
250
NA
<50
<250
500
<10
<100
<500
1,000
<20
<200
<1,000
2,500
<50
<500
<2,500
5,000
<100
<1,000
<5,000
10,000
<200
<2,000
TNTC
25,000
<500
<5,000
TNTC
50,000
<1,000
TNTC
TNTC
250,000
<5,000
TNTC
TNTC
>250,000
TNTC
TNTC
TNTC
Salmonellae Investigations:
The estimation of Enterobacteriaceae cannot be used as a proxy
measurement for presence or absence of Salmonellae species. For
investigations of Salmonellae presence, a standard method Salmonellae
test should be performed from food or from environmental surfaces.
Calibration & Controls:
It is advisable to run positive and negative controls according to Good
Laboratory Practices. Hygiena offers the following controls:
Calibration Control Kit for Hygiena luminometers (Part # PCD4000)
MicroSnap Coliform & E.coli Positive Controls (Part # MS-PC-
COLIFORM)
Storage & Shelf Life:
Store at 2 –8°C. Devices have a shelf life of 12 months. Check expiration
date on label.
Disposal:
Disinfect before disposal. MicroSnap devices can be disinfected by
autoclaving or by soaking in 20% bleach for 1 hour. Then, they can be
placed in the trash. Alternatively, MicroSnap devices may be discarded at a
biohazard waste disposal facility.

INS0193 JAN 2015 Rev A
MicroSnap Enhanced EB Broth –9ml
Directions for use with MicroSnap EB Detection Device (MS2-EB)
Matrix: Liquid Milk Products
Part No: MS1-EB-BROTH-9ML (100)
Safety & Precautions:
MicroSnap Enhanced EB Broth does not pose any health risk when used
correctly. Used devices confirming positive results may be a biohazard and
should be disposed of safely in compliance with Good Laboratory Practice
and Health and Safety Regulations.
1. Enhanced EB Broth tubes are designed for a single use. Do not reuse.
2. Do not use devices after expiration date.
3. Sampling should be done aseptically to avoid cross contamination.
4. Avoid prolonged exposure to light.
5. Ensure proper dilution of sample to be read within the luminometer’s
dynamic range.
6. Ensure proper incubation temperature and time for the test
application.
7. Do not use in the diagnosis of conditions in humans or animals.
User Responsibility:
MicroSnap Enhanced EB Broth does not contain any dyes, therefore, it
alone will not detect bacteria colonies. The incubated broth will contain an
active bacterial culture and should be used with MicroSnap detection
devices. However, the incubated broth contains active viable organisms
that can be subjected to further analysis by any other method. It is the
user’s responsibility to select a test method to evaluate a product by testing
it with sufficient number of samples and microbial challenges to satisfy the
user that the chosen method meets the user’s criteria. Selection of broth,
sampling method, testing protocol, preparation time and handling may
influence recovery and enumeration of specific strains. MicroSnap is a time
dependent test and it is important to follow instructions carefully to achieve
optimal results.
Hygiena Liability:
As with any culturemedium, MicroSnap Enhanced EB Broth results do not
constitute a guarantee of quality of food, beverage products orprocesses that
are testedwith these Devices. Hygiena will not be liable to user or others for
any loss or damage, whether direct or indirect, incidental orconsequential
from use of this Device. If this product is proven to be defective, Hygiena’s
sole obligation will be to replace product, or at its discretion, refundthe
purchase price. Promptly notify Hygiena within 5 days of discovery of any
suspected defect and return product to Hygiena. Please call Customer
Service for a Returned Goods Authorization Number.
Contact information:
If more information is required, please visit us at www.hygiena.com or
contact us at:
Hygiena - Americas
Phone: [phone]
Fax: [phone]
Email: [e-mail]
Hygiena - International
Phone : [phone]
Fax: [phone]
Email: [e-mail]

INS0193 JAN 2015 Rev A
MicroSnap Enhanced EB Broth –9ml
Directions for use with MicroSnap EB Detection Device (MS2-EB)
Matrix: Liquid Milk Products
Part No: MS1-EB-BROTH-9ML (100)
1. Allow milk sample
and broth to equilibrate
to 22 –26 °C. Add 1mL
milk sample directly to
Enhanced EB Broth
vial.
2. Replace cap and
tighten to secure.
3. Shake by hand mixing
or vortexing for 10
seconds.
4. Incubate at 37 ± 0.5 °C
for 6 - 8 hours. Refer to
Table 1 for details.
1. Allow Detection
Device to equilibrate to
room temperature.
Shake to bring liquid in
tube to bottom of tube.
2. Shake by hand
mixing or vortexing for
10 seconds.
3. Aseptically transfer
0.1mL enriched sample
from Enhanced EB Broth
vial to Detection Device.
4. Reassemble device to
original state.
5. Activate device by
breaking Snap-Valve.
Squeeze bulb to
release liquid into tube.
6. After activation,
shake tube gently to
mix sample in liquid.
7. Insert device into
EnSURE and press OK
to initiate measurement.
8. Record results as RLU.
Refer to Table 2 to
interpret results.
8
7
6
5
4
3
2
1
Detection / Measurement
Step: 2
4
3
2
1
Enrichment of Milk Sample
Step: 1
Instructional Video:
www.youtube.com/HygienaTV